CAJ | 학술논문

利用聚合酶链式反应(PCR)技术和基因重组技术克隆了中国明对虾(Fenneropenaeus chinensis)抗脂多糖因子(ALFFc)基因的成熟肽编码序列并构建了pET-DsbA-ALFFc原核表达载体, 转化E.coli BL21 (DE3)后, 经异丙基-β-D-硫代半乳糖苷(IPTG)筛选法获得了1株高表达量重组菌, 并通过实验确定其最佳诱导条件为: 菌液初始OD_(600) 0.7, IPTG终浓度0.8 mmol/L, 培养温度37 ℃, 诱导时间3 h. 重组蛋白经纯化后免疫新西兰大白兔制备了抗血清, 为进一步研究ALFFc的功能奠定了基础.
이용취합매련식반응(PCR)기술화기인중조기술극륭료중국명대하(Fenneropenaeus chinensis)항지다당인자(ALFFc)기인적성숙태편마서렬병구건료pET-DsbA-ALFFc원핵표체재체, 전화E.coli BL21 (DE3)후, 경이병기-β-D-류대반유당감(IPTG)사선법획득료1주고표체량중조균, 병통과실험학정기최가유도조건위: 균액초시OD_(600) 0.7, IPTG종농도0.8 mmol/L, 배양온도37 ℃, 유도시간3 h. 중조단백경순화후면역신서란대백토제비료항혈청, 위진일보연구ALFFc적공능전정료기출.
The sequence coding mature peptide of anti-lipopolysaccharide factor(ALFFc)was cloned from Chinese shrimp Fenneropenaeus chinensis, and a prokaryotic expression vector pET-DsbA-ALFFc was constructed by PCR and recombinant DNA techniques. A high-productivity colony of transformed E. coli BL21(DE3) was obtained by IPTG screening method. The optimal conditions for the introduction of the fused protein were investigated, which are described as follows: final IPTG concentration is 0.8 mmol/L, initial OD_(600) is 0.7, and introduction is conducted at 37 ℃ for 3 h. The purified recombinant protein DsbA-ALFFc was used as antigen to prepare antiserum in New Zealand white rabbit, which provides the basis for further functional analysis of ALFFc.